RNA extraction

Tina anfortin at webnet.qc.ca
Fri Jul 14 20:42:51 EST 2000


Hi everybody!
I have isolated total RNA from non adherent cells with the TRIZOL reagent.
At the end of the procedure, I solubilized the pellet with DEPC-treated
water. After this, I measured OD 260 and 280 and calculated the ratio
260/280. This ratio should be 1.6 to 2.0. Sometimes I have got around 1.6
but generally 1.4 to 1.5  ... it is too low but the RNA is OK on agarose gel
(no apparent degradation). Which step of the Trizol method can I improve to
increase this ratio? I have already read that to calulate the ratio it is
better to take ODs of RNA in TE buffer but other people have already got
good ratios with RNA in water.

Any suggestion?
Thanks a lot!!







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